NZ506024A

NZ506024A - Mixed micellar pharmaceutical delivery system and method of preparation

Info

Publication number: NZ506024A
Application number: NZ506024A
Authority: NZ
Inventors: Pankaj Modi
Original assignee: Generex Pharm Inc
Priority date: 1998-02-10
Filing date: 1999-02-05
Publication date: 2002-02-01
Also published as: DE69937546T2; WO1999040932A9; BR9804295A; PT1053011E; ATE378062T1; DK1053011T3; AU2505399A; DE69937546D1; AU750197B2; US6017545A; EP1053011A1; US6231882B1; WO1999040932A1; ES2296380T3; JP2003522096A; JP4499282B2; EP1053011B1

Abstract

A pharmaceutical composition suitable for delivery through mucosal membranes, and a process for making it. The process comprises a) preparing a pharmaceutical agent composition in micellar form in an aqueous medium which has an alkali metal salicylate in a concentration of from 1 to 10 wt./wt. % of the aqueous micellar pharmaceutical agent composition, an alkali metal C8 to C22 alkyl sulphate in a concentration of from 1 to 10 wt./wt. % of the aqueous micellar pharmaceutical agent composition and a pharmaceutically acceptable edetate in a concentration of from 1 to 10 wt./wt. % of the aqueous micellar pharmaceutical agent composition; b) slowly adding the micellar pharmaceutical agent composition, while mixing, to at least one absorption enhancing compound, while continuing to mix vigorously. The absorption enhancing compounds is selected from the group consisting of saturated phospholipid, unsaturated phospholipid, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linolenic acid, borage oil, evening primrose oil, menthol, trihydroxy oxo cholanylglycine and pharmaceutically acceptable salts of trihydroxy oxo cholarylglycine, glycerin, polyglycerin, lysine, polylysine, polidocanol alkyl ethers and analogues of polidocanol alkyl ethers, triolein and mixtures thereof, wherein the amount of each absorption enhancing compound is present in a concentration of from 1 to 10 wt./wt. % of the total formulation, and the total concentration of alkali metal salicylate, alkali metal C8 to C22 alkyl sulphate, edetate and absorption enhancing compounds is less than 50 wt./wt. % of the formulation.

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number 506024 5002-20399 MTypm MTHETJAR DELIVERY SYSTEM ASTD METHOD OP PREPARATION Field o£ the Invention The present invention relates to an improved 5 delivery system for the administration of large-molecule pharmaceuticals, e-g. peptidic drugs, vaccines and hormones. In particular it relates to pharmaceuticals which may be administered through the oral and nasal membranes. 10 Background to the Invention rn spite of significant efforts in academic and commercial laboratories/ major breakthroughs in oral peptide and protein formulation have not been achieved. Relatively little progress has been made in reaching the IS target of safe and effective oral formulations for peptides and proteins. The major barriers to developing oral formulations for proteins and peptides include poor intrinsic permeability, lumenal and cellular enzymatic degradation, rapid clearance, and chemical stability in 20 the gastrointestinal (GI) tract. Pharmaceutical approaches to address these barriers, which have been successful with traditional small, organic drug molecules, have not readily translated into effective peptide and protein formulations. Although the 25 challenges are significant, the potential therapeutic benefits remain high especially in the field of diabetes treatment using insulin. Scientists have explored various administration routes other than injection for proteins and peptides. 3 0 These routes include oral, intranasal, rectal, vaginal cavities for the effective delivery of large molecules. Out of the above four mentioned routes oral and nasal t AMENDED SHEET — r..v~rm!\ =JO- a- u ; : ao3 276 7687-» +49 89 2t""" 15-02-2000 "uv « •»« CA 009900106 - a - cavities have been of greatest interest to scientists. Both, the oral and nasal membranes offer advantages over other routes of administration. For example, drugs administered through these .membranes have a rapid onset 5 of action, provide therapeutic plasma levels, avoid first pass effect of hepatic metabolism, and avoid exposure of the drug to hostile GI environment. Additional advantages include easy access to the membrane sites so that the drug can be applied, 10 localized and removed easily. Further, there is a good potential for prolonged delivery of large molecules through these membranes. The oral routes have received far atore attention than have the other routes. The sublingual mucosa 15 includes the membrane of ventral surface of the tongue and the floor of the mouth whereas the buccal mucosa constitutes the lining of the cheek. The sublingual mucosa is relatively permeable thus giving rapid absorption and acceptable bioavailability of many drugs. 20 Further, the sublingual mucosa is convenient, acceptable and easily accessible. This route has been investigated clinically for the delivery of a substantial number of drugs. ml»% J x... — n _ i ^ ^ _ j_ . ■*-» i,-AA«s—^f ui. IU^^qwuXBB uv-i crxxx ix UIIC 25 oral mucosa appears to be related to molecular size, lipid solubility and peptide protein ionization. Small molecules, less than 1000 daltons appear to cross mucosa rapidly. Ae molecular size increases, the permeability decreases rapidly. Lipid soluble compounds are more 3 0 permeable than non-lipid soluble molecules. Maximum absorption occurs when molecules are un-ionized or AMENDED SHEET UU v. >/ -r • J . i m : lb- -J.- 0 : 22:52 . id«K u i" 905 276 7687-* . 2;- CA 009900106 - 3 - neutral in electrical charges. Therefore, charged molecules present the biggest challenges to absorption through the oral mucosae. Most proteinic drug molecules are extremely large 5 molecules with molecular weights exceeding 6000 daltons. These large molecules have very poor lipid solubility and are practically impermeable. Substances that facilitate the absorption or transport of large molecules (>2000 daltons) across biological membranes 0 are known as enhancers, (Lee et al., Critical Reviews in Therapeutic drug Carrier Systems-, 8, 91, 1991; Lee et al., Critical Reviews in Therapeutic drug Carrier Systems, 8, 115, 1991, 1992). Enhancers may be characterized as chelators, bile salts, fatty acids, 15 synthetic hydrophilic and hydrophobic compounds, and biodegradable polymeric compounds. Various mechanisms of action of enhancers have been proposed. These mechanisms of action, at least far protein and peptidic drugs, include (1) reducing 2 0 viscosity and/or elasticity of the mucous layer, (2) facilitating transcellular transport by increasing the fluidity of the lipid bilayer of membranes, and (3) increasing the thermodynamic activity of drugs (Critical Rev. 117-1 gq. 1391. 1,99-2}-: - 25 Many enhancers have been tested so far and some have been found to be effective in facilitating mucosal administration of large molecule drugs. However, hardly any penetration enhancing products have reached the marketplace. Reasons for this include the lack of a 30 satisfactory safety profile respecting irritation, lowering of the barrier function, and impairment of the AMENDED SHEET NCHILM 02 U T - U . I IP .3 5- 2- u : 22:53 _ tup i umi u iiivu1- 905 276 7687-. „«^9. ?? CA 009900106 - 4 - raucocilliary clearance protective mechanism. The main factor to be considered in the use of enhancers, especially bile salts and some protein solubilizing agents, is extremely bitter and unpleasant taste. This 5 makes their UBe almost impossible for human consumption on a daily basis. Several approaches were utilised to improve the taste of the bile salts based delivery systems, but none of them are commercially acceptable for human consumption to date., The approaches utilized 10 include patches for buccal mucosa, bilayer tablets, controlled release tablets/ use of protease inhibitors, buccally administered film patch devices, and various polymer matrices. The basic problem associated with the above 15 technologies is the use of large quantities of bila acids and their salts to promote the transport of large molecules through membranes in the form of localized delivery systems using patches or tablets. In spite of using protease inhibitors and polymer coatings, the 20 technologies failed to deliver proteinic drugs in the required therapeutic concentrations. Further, the problem is compounded because of the localized site effect of the patch, which resulted in severe tissue iuCuuiuL«~nwtfW O uu 25 large molecules via the oral, nasal, rectal, and vaginal routes using single bile acids or enhancing agents in combination with protease inhibitors and biodegradable polymeric materials. However, it is extremely difficult to achieve therapeutic levels of proteinic drugs using 3 0 these formulations. Single enhancing agents fail to loosen tight cellular junctions in the oral, nasal, AMENDED SHEET ENCHEN 02 : 15- 2- 0 : 22:53 : 905 276 7687-* +49 89 nnnnn/nne 02-2000-T'uutit. uniMi . unu u invju nvmvv oAUOyyUUlUo - 5 - rectal and vaginal cavities for a required period of time to allow the passage of large molecules through the mucosal membranes without further degradation. This problem makes it impractical to use the above mentioned 5 systems for a commercial purpose. In order to overcome the above mentioned problem of the bitter taste, irritation and the penetration of large molecules through the sublingual, buccal and GI tract mucosal lining, a system has now been designed 10 where protein drug Is encapsulated in mixed micelles made up of a combination of enhancers, e.g. yolk proteins (lecithins). This system allows opening of the paracellular j"unctions (tight junctions) in oral membranes as well as in the GI tract by gl motility 15 movement with a high degree of protease activity preserved, and protects molecules from premature degradation in the hostile acidic and proteolytic GI environment. It is believed that the mixed micelles encapsulate 2 0 molecules with a high degree of efficiency {>90% encapsulation) . These mixed micelles are extremely small in size (1 nm to 10 run) , and are smaller than the pores of the membranes in the oral cavity or the GI TJm J ** 4_^. — T-^ —. "1 J —-. -3 ■*_! J- 1 „ —j.o—w-L^JL.ClUi.C3 UCXXCVCU U4AC3.U UlltT 25 small size of mixed micelles helps encapsulated molecules penetrate efficiently through the mucosal membranes of the oral cavity. The absorption of proteins and peptides is believed to be enhanced by the diffusion of large molecules 3 0 entrapped in the mixed micellar form through the aqueous pores and the cell structure perturbation of the tight l AMENDED SHEET * OH : JS- '2- 0 : 122 • 53 b05 *'7(1 7L>£J7-» +4-9 P.H _ , 5-02-2000'MJU unnn.vjn- v .tv^u ~ ,v t„a, WB„J — CA 009900106 paracellular junctions. The amount of physiologically active peptide or protein in the compositions of this invention is typically a quantity tbat provides an effective amount 5 of the drug to produce the physiological activity (therapeutic plasma level) for which the peptide or protein is being administered. In consideration of the fact that the bioavailability of any active substance can never be 100%, that is to say the administered dose 10 of the active drug is not completely absorbed, it is preferable to incorporate a slightly larger amount than the desired dosage. Where the dosage form is a spray (aerosol) or the like which is repeatedly dispensed from cbe same container, it' is recommended that the unit dose 15 will be slightly greater than the desired dose. It should be understood that dosage should vary with species of warm blooded animals such as man, domestic animals, and their body weights. The composition of thia invention, is prepared as jnicrofine droplets (1 to 2 0 10 nm or less) by virtue of the preparation methods and the characteristics of suitable combinations of enhancer compounds used. Atomizer or aerosol spray devices (metered dose inhalers or nebulizers) may be 'useful in eff«ciri^xg~a~sufficient~r^duc:irion in particle size "for 25 effective inhalation from the nasal or oral cavity so that the drug may be successfully absorbed or reach the specific site. The therapeutic composition of the present invention can be stored at room temperature or at cold 30 temperatures. It is preferable to store proteinic drugs at cold temperatures to prevent their degradation and to AMENDED SHEET - fci\iC_HbN oii : 15- 2- 0 : 22*5+ : 905 276 7687-r -t49 83 22 _ . -02-2000L.nnmu'-.n U rovviu mb'Vuij 'CA 009900106 - 7 - extend their shelf life. The mixed raicellar therapeutic composition of the invention is applied to the mucosal membranes/ and the sites of administration may be the same as those used for usual mucosal therapeutic 5 preparations. Generally, oral, transdermal and nasal sites are favoured, but the composition, can be applied to the rectal and vaginal mucosa. According to the physiologically active, peptide or protein used, the dosage form and the site of administration, a specific 10 administration method can be selected. The term "edetate" is used herein to refer to pharmaceutically acceptable salts of ethylenediaminetetraacetic acid. It has alBO been found that improvements in 15 penetration and absorption of mixed micellar formulations can be achieved by mixing the mixed micellar formulation with propellants such as tetrafluoroethane, heptafluoroethane, dimethylfluoropropane, tetrafluoropropane, butane, 20 isobutane, dimethyl ether and other non-CFC and CFC propellants. Preferably they are delivered through metered dose spray devices. Metered dose inhalers are known and are a popular pulmonary drug delivery form for . RjrYTTV^_. T<ho_r< r-fl o on f_ f ^ ^— wmO 25 propellant is intended to improve the quality of absorption, stability and performance of many formulations. The compositions have been selected to give enhancement in the penetration through pores, and facilitate absorption of the drugs to reach therapeutic 30 levels in the plasma. The present formulation may be absorbed buccally, by ensuring that the person does not AMENDED SHEET fcNCHLN 02 :15- -j.- o : 22:54 : 905 276 7687-> +49 89 22"" -02-2000uuu u irv\j\i CA009900106 - 8 - inhale the formulation as it is sprayed- One of the other benefits of using an atomizer or inhaler is tbat the potential for contamination is minimized because the devices are self contained. 5 Summary of the Invention Accordingly, the present invention provides a mixed micellar pharmaceutical formulation comprising a proteinic pharmaceutical agent in micellar form, water, an alkali metal lauryl sulphate in a concentration of 10 from 1 to 10 wt./wt.% of the total formulation, a pharmaceutically acceptable edetate in a concentration of from 1 to 10 wt./wt.% of the total formulation, at least one alkali metal salicylate in a concentration, of from 1 to 10 wt./wt,.% of the total formulation, and at 15 least one absorption enhancing compound selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenaxypolyetboxyethanol, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, 20 linolenic acid, borage oil, evening of primrose oil, trihydroxy oxo cholanylglycine, glycerin, polyglycexin, lysine, polylysine, triolein and mixtures thereof, wherein the amount of each absorption enhancing compound -1 4 n m WJIW 4 AM +» 1 ft »kf+» / *.*4* V _ ™ —" ■»»-#*, Wr V IA u'j. W41 Wte WAII VV *¥ ■* • ~f »» w f 9 25 of the total formulation, and the total concentration of absorption enhancing compounds is less than 50 wt./wt.% of the formulation. In an embodiment, the alkali metal lauryl sulphate, the edetate and the alkali metal salicylate are each in 3 0 a concentration of from 2 to 5 wt./wt-* of the total 1 AMENDED SHEET fclNCHHN 02 :iB- 2- 0 : 22-54 : 90S 27f» 7687- +49 89 2C ~» ooonnnirw 15-02-2000uyv ~ * j ■ * u III VKU iiu'Divu CA 009900106 - 9 - formulation. In one embodiment/ the edetate is an alkali metal edetate. Preferably the alkali metal edetate is Selected from the group consisting of disodium edetate, 5 dipotassium edetate, and combinations thereof. In another embodiment, the alkali metal lauryl sulphate is sodium lauryl sulphate. In a further embodiment*, the alkali metal salicylate is sodium salicylate. 10 In another embodiment, the lecithin is selected from the group consisting of saturated phospholipid, e.g. Phospholipon-H {trade mark) saturated phospholipid, unsaturated phospholipid, e.g. Phospholipon-G (trade mark) unsaturated phospholipid, phosphatidylcholine, 15 phosphatidyl serine, sphingomyelin, phosphat idyl ethanol amine, cephalin, and lysolecithin. In one embodiment, one of the absorption enhancing compounds is selected from the group consisting of hyaluronic acid, pharmaceutically acceptable salts of 20 hyaluronic acid and mixtures thereof, the concentration of such absorption enhancing compound being from about 1 to about 5 wt./wt.%- In another embodiment, suitable for delivery iita.»ax"p£a.»fci«a.yes, mixeu uu.~cej.xar prLaxma.ceutxca.1. 25 formulation is suitably diluted to avoid irritation of the nasal passages. Another aspect of the present invention provides a mixed micellar pharmaceutical formulation suitable for delivery by means of an aerosol and comprising a 30 pharmaceutical agent in micellar form, water, an alkali metal C8 to C22 alkyl sulphate in a concentration of ' AMENDED SHEET y tiNCHbTvi 02 15-02-2000J y 'J :15- 2- 0 ; 22-54 jnn.uunii u iyivu-. 305 !2TG 7GS7- +4.Q PQ .i«• «>T — CA 009900106 - 10 - from 1 to 10 wt./wt.% of the total formulation, a pbarioaceutically acceptable edetate in a concentration of from l to 10 wt./wt.% of the total formulation, at least one alkali metal salicylate in a concentration of 5 front 1 to 10 wt./wt<% of the total formulation, and at least one absorption enhancing compound selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic acid, lactic 10 acid, chamomile extract, cucumber extract, oleic acid, linolenic acid, borage oil, evening primrose oily menthol, trihydroxy oxo cholanylglycine and pharmaceutically acceptable salts thereof, glycerin,, polyglycerin, lysine, polylysine, polidocanol alkyl 15 ethers and analogues thereof, triolein and mixtures thereof, wherein each absorption enhancing compound is present in a concentration of from 1 to 10 wt. /wt. %■ of the total formulation, and the total concentration of absorption enhancing compounds is less than 50 wt./wt. % 20 of the formulation. Yet another aspect of the present invention provides that the mixed micellar aerosol pharmaceutical formulation additionally comprises i) a phenol selected f rervthe group-—consisting—of—ph®nol lustiiyl—phenol in_ 25 a concentration of from 1 to 10 wt./wt.% of the total formulation, and i) a propellant selected from the group consisting of C1-C2 dialkyl ether, butanes, fluorocarbon propel lant, hydrogen-containing fluorocarbon propel lant, chlorofluorocarbon propellant, hydrogen-containing 3 0 chlorofluorocarbon propellant, and mixtures thereof. In one embodiment, the alkali metal C8 to C22 alkyl AMENDED SHEET E\CHE\ 02 :15- 2- 0 : 22:55 : 905 276 7687-* +4-9 89 2.3 " 15-02-2000 ^v -r,u-"l« unmwsan.. u uvuu . v» j CA009900106 - 11 - sulphate is in a concentration of from 2 to 5 wt./wfc.% of the total formulation. In another embodiment, the alkali metal C8 to C22 alkyl sulphate is sodium lauryl sulphate. 5 In another embodiment, the lecithin is saturated or unsaturated, preferably selected from the group consisting of phosphatidylcholine, phosphatidyl serine, sphingomyelin, phosphatidylethanolamine, cephalin, and lysolecithin. 10 In yet another embodiment r one of the absorption enhancing compounds is selected from the group consisting of hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, polidocanol alkyl ethers, trihydroxy oxo cholanyl glycine, polyoxyethylene 15 ethers and mixtures thereof, the concentration of such absorption enhancing compound being from about 1 to about 5 wt./wt.%. In yet another embodiment, the formulation comprises a combination selected from the group 2 0 consisting of i) sodium lauryl sulphate, sodium salicylate, disodium edetate, saturated phospolipid and sodium hyaluronate; ii) sodium lauryl sulphate, sodium salicylate, disodium edetate, lecithin and sodium ~ ' byalur dha tel ~Xi~i" }^sodium"ratii^l~s1IIphate^"sodim 25 salicylate, disodium edetate, sodium hyaluronate and evening primrose oil; iv) sodium lauryl sulphate, sodium salicylate, disodium edetate, saturated phospolipid and bacitracin; v) sodium lauryl sulphate, sodium salicylate, disodium edetate, saturated phospholipid, 30 sodium hyaluronate, and bacitracin; vi) sodium lauryl sulphate, sodium salicylate, disodium edetate, sodium t AMENDED SHEET t fc-NCHHV 02 =15- 2- 0 : 22 = 55 : 905 276 7687-* +49 89 on------ 15-02-2000 J"'j 1 u"vu « Ii-vw „u.v WJ CA 009900106 - 12 - hyaluronate, oleic acid and gamma linoleic acid; vii) sodium lauryl sulphate, sodium salicylate, disodium edetate, saturated phospholipid, and glycolic acid; and. viii) sodium lauryl sulphate, sodium solicylate, 5 disodium edetate, saturated phospholipid, glycolic acid and lactid acid. Preferably, the ratio of pharmaceutical agent, e.g. insulin, to propellant is from 1:19 to 1:3. In another embodiment, the propellant is selected 10 from the group consisting of tetrafluoroethane, tetraf luoropropane, dimethyl f luoropropane, heptafluoropropane, dimethyl ether, n-butane and isobuuane. In yet another embodiment, the mixed micellar 15 pharmaceutical formulation is contained in an aerosol dispenser. For insulin-containing and some other formulations, the formulation may also contain at least one inorganic salt which opens channels in the gastrointestinal tract 20 and may provide additional stimulation to release insulin. Non-limiting examples of inorganic salts are bodium, potassium, calcium and zinc salts, especially sodium chloride, potassium chloride, calcium chloride, .. —,-i.— — — t- n J -a ^ _ «a j . * i?WXUliri-»iUUiW44Wk»"Wl 25 It will be recognized by those skilled in the art that for many pharmaceutical formulations it is usual to add at least one antioxidant to prevent degradation and oxidation of the pharmaceutically active ingredients. It will also be understood by those skilled in the art 3 0 that colorants, flavouring agents and non-therapeutic amounts of other compounds may be included in the formulation. Typical flavouring agents are menthol and AMENDED SHEET - KNCHU.M 02 :I5- 2- 0 : 22:55 : 905 27t5 7C87- +4-fi 83 2:i _ . -02-2000UIJU „niuvuniv <1 ,vvn ny.«i NJ CA 009900106 - 13 - sorbitol- In one embodiment the antioxidant is selected from the group consisting of tocopherol, deteroxime mesylate, methyl paraben, ethyl paraben and ascorbic acid and 5 mixtures thereof. A preferred antioxidant is tocopherol. In a preferred embodiment, at least one protease inhibitor is added to the formulation to inhibit degradation of the pharmaceutical agent by the action of 10 proteolytic enzymes. Of the known protease inhibitors, most are effective at concentrations of from 1 to 3 wt./wt.% of the formulation* Non-limiting examples of effective protease inhibitors are bacitracin, soyabean trypsin, aprotinin 15 and bacitracin derivatives, e.g. bacitracin methylene disalicylate. Bacitracin is the most effective of those named when used in concentrations of frem 1.5 to 2 wt, /wt. % . Soyabean trypsin and aprofcinin may be used in concentrations of about 1 to 2 wt. /wt - % of the 20 formulation. The formulation suitable for delivery through oral mucosal membranes may be in chewable form, in which case it will be necessary to add ingredients suitable for bu Q i"i™£ SuC,ii~ iiiyX cdi tS irliOivUics—gUS. X— gvwTiT p ovri-sr sd. 25 acacia, carrageenin, beeswax and xanthan gum. The pharmaceutical agent may be selected from a wide variety of macroiao 1 ecular agents, depending on the disorder being treated, generally with molecular weights greater than about 1000 and especially between about 30 1000 and 2 000 000. Preferred pharmaceutical agents are selected from the group consisting of insulin, heparin. AMENDED SHEET " "" ~Z~™'ENCHEN 02 :15- 2" 0 : 22:55 : SOB 276 7687- +4-9 89 2£ ~ . nnnnnmAe 15-02-2000 u u ,J un.vii lunn u mvuu ivimjj CA 009900106 - 14 - low molecular weight heparin, hirulog, hirugen, huridine, interferons, interleukins, cytokins, mono and polyclonal antibodies, immunoglobins, chemofcherapeutic agents, vaccines, glycoproteins, bacterial toxoids, 5 hormones, calcitonins, insulin like growth factors (IGF), glucagon like peptides (GLP-1), large molecule antibiotics, protein based thrombolytic compounds, platelet inhibitors, DNA, RNA, gene therapeutics and arxtisense oligonucleotides, and small molecule drugs, 10 e.g. opioids, narcotics, analgesics, NSAIDS, steroids, hypnotics, pharmaceutical preparations for the treatment of chronic pain, morphine and the like. The present invention also provides a process for making a pharmaceutical formulation suitable for 15 delivery through transdermal membranes comprising: a) preparing a proteinic pharmaceutical agent formulation in micellar form in an aqueous medium which has an alkali metal salicylate in. a concentration of from 1 to 10 wt«/wt.% of the aqueous micellar 20 pharmaceutical agent formulation, an alkali metal lauryl sulphate in a concentration of from 1 to 10 wt./wt.% of the aqueous micellar pharmaceutical agent formulation and a pharmaceutically acceptable edetate in a COnCSIit 2T2.W i OH— Of SwOJTt— 1* tO—i 0 ^ t 25 micellar pharmaceutical agent formulation; b) slowly adding the micellar proteinic pharmaceutical agent composition to at least one of the absorption enhancing compounds selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically 3 0 acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic acid, lactic AMENDED SHEET } •- tM.hCM : lb- J.- u • ^2-50 : aU5 27G 76C7-» +4-9 «9 20 15-02-2000""" nv.Mi-u CA009900106 - IS - acid, chamomile extract, cucumber extract, oleic acid, linolenic acid, borage oil, evening of primrose oil, trihydroxy oxo cholanylglycine, glycerin, polyglycerin, lysine, polylysine, triolein and mixtures thereof, while 5 mixing vigorously, to form a mixed micellar formulation; wherein the absorption enhancing compounds are each present in a concentration of from 1 to 10 wt./wt,% of the total formulation, and the total concentration of alkali metal salicylate, alkali metal lauryl sulphate, 10 edetate and absorption enhancing compounds is less than 50 vrt./wt.% of the formulation. In one embodiment, the process provides an additional step of adding, while continuing vigorous mixing, at least one absorption enhancing compound 15 different from that added in step b), selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, 20 linolenic acid, borage oil, evening of primrose oil, trihydroxy oxo cholanylglycine, glycerin, polyglycerin, lysine, poly lysine, triolein and mixtures thereof. In one embodiment the alkali metal lauryl sulphate _ t_ e erv-1 -i urn 1 anrvl en 1 rihate . 25 In another embodiment the alkali metal salicylate is sodium salicylate. In a further embodiment the alkali metal edetate may be selected from the group consisting of disodium edetate and dipotassium edetate. 3 0 In yet another embodiment, the formulation has a combination selected from the group consisting of AMENDED SHEET - Uli : 15- 2- 0 : 22:56 : 1305 270 7687^ +43 89 2C 0 « nnnnnnmc 15 02-2000 '"J U lr "niiiiun.i 1/ mvu«. uvv/i-ry CA0099UUlUb - 16 - i) saturated phospholipid ana sodium hyaluronate; ii) saturated phospholipid and glycolic acid; iii) lecithin and sodium hyaluronate; iv) saturated phospholipid, glycolic acid and lactic acid; v) sodium hyaluronate and 5 evening primrose oil; vi) saturated phospholipid and bacitracin; vii) saturated phospholipid, sodium hyaluronate and bacitracin? and viii) sodium hyaluronate, oleic acid, and gamma linoleic acid. The present invention also provides a process for 10 making a mixed mi eel lax pharmaceutical formulation suitable for delivery by means of an aerosol comprising: a) preparing a pharmaceutical agent formulation in micellar form in an aqueous medium which has an alkali metal C8 to C22 alkyl sulphate in a concentration of 15 from l to 10 wt./wt,% of the aqueous micellar pharmaceutical agent formulation, a pharmaceutically acceptable edetate in a concentxatiori. of from 1 to 10 wt./wt.% of the aqueous micellar pharmaceutical agent formulation at least one alkali metal salicylate in a 20 concentration of from 1 to 10 wt./wt.% of the aqueous micellar pharmaceutical agent formulation; b) slowly adding the micellar proteinic pharmaceutical agent formulation to at least one absorption enhancing r« r*vrrmyyi t t> ^ oal a f v/xifj r-» -? f- 25 hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linolenic acid, borage oil, evening primrose oil, menthol, trihydroxy oxo cholanylglycine and 30 pharmaceutically acceptable salts thereof, glycerin, polyglycerin, lysine, poly lysine, polidocanol alkyl AMENDED SHEET . 'i. _ >■>-».... n - i o - -a. - u , .jji • /to / /—* 4-4H * -02-2000" '*' U ~"J-'"' Ufim.on.. u »vUk n«.viTU CA 009900106 - 17 - ethers and analogues thereof, triolein and mixtures thereof, while mixing vigorously, to form a mixed micellar composition; and optionally c) an additional step of adding, while continuing 5 vigorous mixing, at least one absorption enhancing compound different from that added in step b) , selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber 10 extract, oleic acid," linoleic acid, linolenic acid, monoolein, borage oil, evening primrose oil, glycerin, polyglycerin, lysine, polylysine, triolein, polyoxyethylene ethers and analogues thereof, polidocanol alkyl ethers and analogues thereof, and 15 mixtures thereof; d) mixing the mixed micellar composition resulting from steps a) to c) with a phenol selected from the group consisting of phenol, m-cresol and mixtures thereof; and subsequently 20 e) placing the formulation into an aerosol dispenser and charging the dispenser with a propellant; wherein each of the absorption enhancing compounds are present in a concentration of from 1 to 10 wt./wt.% ^ Hp +• Is n I- 1 il ^ J *■» *■ ^ ^ •*"— * VMXAt* W4AC UWUttX V" WA. 25 alkali metal salicylate, alkali metal C8 to C22 alkyl sulphate, edetate and absorption enhancing compounds is less than 50 wt./wt.% of the formulation. The vigorous mixing may be accomplished by using high speed stirrers, e.g. magnetic stirrers or propellor 30 stirrers, or by sonication, In one embodiment, the mixed micellar formulation AMENDED SHEET -"JENCHEN 02 -15- 2- 0 : 22:57 ; 905 276 76SS7-* +49 89 2:,-,, »nnQnn1 ne 15-02-2000 u u u ^ ^ bn»i. •u'\u u »vu\, UA uuyyuuiuo - 18 - is formed fay sonication of the aqueous micellar pharmaceutical agent composition in the presence of lecithin. Detailed Description of Preferred Brrihod-inwant-.s 5 The present invention provides an improved method for delivery of macromolecular (high molecular weight} pharmaceutical agents, particularly through the membranes in the nose, mouth, vagina or rectum. The preferred delivery is through oral and nasal cavities. 10 The pharmaceutical agents cover a wide spectrum of agents, including proteins, peptides, hormones, vaccines and drugs. The molecular weights of the macromolecular pharmaceutical agents are preferably above 1000, especially between 1000 and 2 000 000. 15 For example, hormones which may be administered with the present invention include thyroids, androgens, estrogens, prostaglandins, somatotropins, gonadotropins, erythropoetin, interferons, interleukins, steroids arid cytokins. Vaccines which may be administered with the 20 present invention include bacterial and viral vaccines such as vaccines for hepatitis, influenza, tuberculosis, canary pox, chicken pox, measleB, mumps, rubella, pneumonia, BCG, HIV and AIDS. Bacterial toxoids which - iuay cL<ifuiiXXSc c£ ci ua xliypx ScicHt ialVSIit i On'~3.nduds 25 diphtheria, tetanus, pseudonomas and mycobactrium tuberculosis. Examples of specific cardiovascular or thrombolytic agents include heparin, hirugen, hirulos and hirudin. Large molecules usefully administered with the present invention include monoclonal antibodies, 3 0 polyclonal antibodies and iromunoglobins. As will be understood, the concentration of the AMENDED SHEET 02 :15- 2- 0 22 = 57 : 905 276 7687- +49 89 2(pA nnQQnnmfi 02 2000 U " u ' ^1,1 unniuunii u ,»iuuu OA UUSSUU IVJD - IS - pharmaceutical agent is an amount sufficient to be effective in treating or preventing a disorder or to regulate a physiological condition in an animal or human. The concentration or amount of pharmaceutical 5 agent administered will depend on the parameters determined for the agent and the method of administration, e.g. oral, nasal. For example, nasal formulations tend to require much lower concentrations of some ingredients in order to avoid irritation or 10 burning of the nasal passages. It is sometimes desirable to dilute an oral formulation up to 10-100 times in order to provide a suitable nasal formulation. The mixed micellar formulation is prepared by first preparing a first micellar composition which contains 15 the pharmaceutically active agents, alkali metal C8 to C22 alkyl sulphate, edetate and alkali metal salicylate. For those compositions intended for administration through the nasal, oral, vaginal or rectal cavities, the first micellar composition is then added to at least one 20 of the absorption enhancing compounds to form a mixed micellar composition. At least one other absorption enhancing compound may also be added subsequently. Preferably the first absorption enhancing compound is ~ — - — ■ — = ■ ■ — - — ~ ■ 25 When making the aerosol formulation, the phenol and/or m-cresol and/or isotonic agent are then added. The formulation is then put into an aerosol dispenser and the dispenser charged with propellant. The preferred propellants are hydrogen-containing 30 chlorofluorocarbons, hydrogen-containing fluorocarbons, dimethyl ether and diethyl ether. Even more preferred AMENDED SHEET - e.v~niii\ (J2 : 1&- 12- 0 : 22:57 : 905 276 7bS7-» +49 89 22^. „ 15-02-2000 UUU unmi.umi u nvgi-v CA009900106 - 20 - is hydrofluoroalkane (HFA) 134a {1,1,1,2 tetraf luoroethane). Although the present invention has such, wide applicability, the invention is described hereinafter 5 with particular reference to insulin and its analogues, which are used for the treatment of diabetes. As indicated hereinbefore, the compositions of the present invention require that the pharmaceutical formulation be in mixed, micellar form. t 10 in the case of insulin, which is intended for administration through nasal or oral cavities, the first micellar solution may be made by adding a buffer solution to powdered insulin, and then stirring until the powder is dissolved and a clear solution is 15 obtained. A typical buffer solution is an aqueous solution of sodium salicylate and sodium lauryl sulphate and disodium edetate. Typical concentration of sodium salicylate and sodium lauryl sulphate in the aqueous solution are about 1 to 10 wt./wt.% of each compound in 2 0 the solution. Typically, insulin is present in the micellar solution in an amount which will give a concentration of about 2 to 4 wt./wt.% of the final formulation. Typically the concentration may be about xu~ wcT/ wi, ^~~ot clis Iirst mxceixar composicion. ~ 25 The micellar solution is then added slowly to the first absorption enhancing compound, e.g. lecithin while mixing vigorously, e.g. sonicating, to form a mixed micelle liposomal solution. At least one other absorption enhancing compound selected from the group 3 0 consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, AMENDED SHEET • "" LNCHbN 02 : 15- 2- 0 : 22:58 • 905 27U 7687-* +49 8y 25 n A nooonninc 15-02-2000u u ^ y •v'1 i-'nw: uM u mvjm UAOOyyUUiUb - 21 - octylphenoxypolysthoxyethanol, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, liaolenic acid, borage oil, evening primrose oil, trihydroxy oxo cholanylglycine, glycerin, polyglycerin, 5 lysine, polylysine, triolein is then added. The mixing may be done with a high speed mixer or sonicator to ensure uniform micelle particle size distribution within the formulation. Each of the absorption enhancing compounds, when 10 present, is in a concentration of from 1, to 10 wt./wt.% of the total formulation. Preferred salts of hyaluronic acid are alkali metal hyaluronates, alkaline earth hyaluronates and aluminium hyaluronate. The preferred salt is sodium hyaluronate. 15 The preferred concentration of hyaluronic acid or pharmaceutically acceptable salts of hyaluronic acid is from l to 5 wt./wt.% of the total formulation. An even more preferred range is from 1.5 to 3.5 wt. /wt. % of the total formulation. 20 Other ingredients may be added to the mixed micellar solution. For example, flavouring agents, antioxidants, salts, protease inhibitors or other pharmaceutically acceptable compounds may be added. . MM A ^ i" -«■ *m. #1 mm ■»*» C +• X* « mmm •! « T n mi "5 1 4 xii cucx ax / Luc oX'ac v L V'c^ ^ j.c»ia xn 25 the solution is about 1 to 10 nm, and preferably from 1 to S nm. Such a size distribution ensures effective absorption of the formulation, and therefore the pharmaceutical agent, through the membranes, for example the membranes in the oral and nasal cavities. 30 The specific concentrations of the essential ingredients can be determined by relatively AMENDED SHEET U.i = LO- n- v • ^: 5ti : &05 276 7687-» +49 89 2. _ . r\n 15-02-2000 wtu 11 un"ll,u™1 u lwvii-j CA009900106 - 22 - straightforward experimentation. For absorption through tiie nasal and oral cavities, it is often desirable to increase, e.g. double or triple, the dosage which is normally required through injection or administration 5 through the gastrointestinal tract. As will be understood, the amount of each, component of the formulation will vary depending on the pharmaceutical agent and the site of application. Preferred formulations for oral or nasal application 10 have the following combinations: i) sodium lauryl sulphate, sodium salicylate, disodium edetate, Phospholipon-H and sodium hyaluronate; ii) sodium lauryl sulphate, sodium salicylate, disodium edetate, lecithin and sodium hyaluronate; iii) sodium lauryl sulphate, 15 sodium salicylate, disodium edetate, sodium hyaluronate and evening primrose oil; iv) sodium lauryl sulphate, sodium salicylate, disodium edetate, Phospholipon-H and bacitracin; v) sodium lauryl sulphate, sodium salicylate, disodium edetate, Phospholipon-H, sodium 20 hyaluronate and bacitracin; and vi) sodium lauryl sulphate, sodium salicylate, disodium edetate, sodium hyaluronate, oleic acid and gamma linoleic acid. For aerosol formulations, the addition of a mixture Ox" yjiesiiux ^iaa'm^cresoi' is'^preierrea"; sucn~air aerosol 25 formulation may then be placed in an aerosol dispenser and then charged with a propellant, preferably a non-CFC propellant. The therapeutic compositions of the present invention may be stored at room temperature or at a cold 30 temperature. Storage of proteinic drugs is preferable at a cold temperature to prevent degradation of the ' AMENDED SHEET - 23 - drugs and to extend their shelf life. As indicated hereinbefore, generally, oral and nasal membranes are the favourite sites of the administration but the composition can be applied to the 5 rectal and vaginal mucosa. According to the physiologically active peptide or protein used, the dosage form, and the site of administration, a specific administration, method can be selected. The formulation of this invention is generally 10 prepared as microfine mixed micellar particles (1 to 10 ma or less) by virtue of the preparation methods and characteristics of suitable combinations of absorption enhancers used. For oral and nasal application, sprays are 15 preferable, but drops, chewable tablets, chewable gum and other suitable forms may also be used. Utilization of atomizer or aerosol spray devices (metered dose inhalers or nebulizers) can be used to further reduce the particle size for effective inhalation from the 20 nasal or oral cavity so the drug may successfully reach the specific site and be absorbed. It is also possible to'utilize a drug delivery system such that an enteric coating is applied to the gelatin capsule to cause the ^T ^ A- _ l i _ 3 1 ~ A.!*-. — J — j- i W.SS a, U.J.O. UUUUC1AU1U UAXCS 25 proximity of the large intestine and not in the stomach. The invention is illustrated by reference to the following examples. Example 1 A first experiment was conducted to provide data 30 for comparative purposes. This example does not fall within the scope of the present invention. AMENDED SHEET 'Nv_rto\i u_: • ia- li- o : lili' Ob : tlUb "J.7G 7(J87-» +4-i) 89 2.H "* ' 15-02-2000J'JU u MV«itj 1CA009900106 - 24 - A solution was prepared using 0.5 g sodium lauryl sulphate, 0-5 g sodium salicylate and 0.25 g disodium edetate dissolved in 10 mL of water. To this solution 40 mg (1000 units) of insulin was added and dissolved S completely while stirring, to give about 100 unite/mL insulin solution. In one set of tests, five healthy non-diabetic human volunteers were tested with insulin, by injection. In another set of tests the volunteers were tested with. 10 insulin, taken orally. The volunteers fasted from midnight prior to the test, with no food being taken during the 4 hour study. On the first day, the volunteers received 10 units insulin by injection (regular fast acting insulin, 15 available from Eli Lilly) . On the second day, the volunteers received 100 units (l mL volume per drop, approximately 20 drops) of the above-prepared oral insulin (10 times the injection dose). In both tests, blood glucose levels were monitored every 15 minutes by 20 Bayer's Gluccaneter Elite. The average results for the five volunteers, of the first day's trial (sub-cutaneous injection with 10 units) were as followsi 25 Time*: 0 15 30 60 75 90 120 150 180 Avg: 5.8 5.8 5.4 5.0 4.6 4.3 3.8 3.6 3.4 Time*; 210 24 0 Avg: 4.2 4.5 * time in minutes 3 0 The results for each of the five volunteers, of the second day's trial (oral drops with 100 units) were as AMENDED SHEET ux : 15- 2- 0 : 22-59 : 005 27<o 7087-^ +49 89 2C 5-02-2000u '*v yni» .T \» u' u n v • v/1 T J CA 009900106 - 25 - follows Table II Time*: 0 15 30 60 75 90 120 150 180 Subject NOS: 1 6.2 5.8 5.2 5.0 4.9 5.0 5,0 4.8 4.7 2 5.8 5.4 5.0 4.7 4.9 4.3 5.0 5.5 5.2 3 4.8 4.6 4.3 4.3 4.4 4.6 4.8 4.7 5.2 4 6.6 6.1 5.8 5.5 5.1 4.9 5.0 5.0 5.9 5 6.0 5.8 5.7 5.5 5.1 4.8 4.7 4.9 5.0 Time*: 210 240 Subject NOS: 1 5.5 6.0 2 5.8 6.1 3 5.5 5.1 4 6.2 6.8 5 5,9 6.7 * time in minutes These tests indicate that compared to the injection method, oral insulin gives a faster onset of action and 2 0 lowers blood glucose levels without creating hypoglycaeiaic condition. Due to the hepatic glucose production, there was a rebound effect. This is believed to be due to the incomplete absorption of insulin. 2"-" " 25 Another experiment, not within the scope of the present invention, was performed for comparative purposes. Oral insulin (100 units) was formulated in (Phospholipon-H, 10 mg) without any sodium lauryl 30 sulphate, sodium, salicylate, edetate or absorption enhancers, to evaluate its efficacy of blood glucose AMENDED SHEET ir -• 'J2 :15- 2- () ; 22-59 : 905 27f-i 7687-» +4.0 00 O; _ -| 5-Q2-2000 W "*U unnn.umv u nvuirii CA 009900106 - 26 - lowering in a fasted state, for healthy volunteers. Volunteers were asked to fast overnight and not have any breakfast prior to dosing. Volunteers were asked to take this oral insulin formulation, in their 5 mouth and swallow it. Blood glucose levels were monitored every 15 minutes using Bayer's glucometer Elite for 3 hours, and the average results for 5 volunteers are shown in Table III. Table III 10 Time*: 0 IS 30 45 60 75 90 120 150 180 Avg: 5.6 5.8 5.8 5.7 5.7 5.8 5.7 5.7 5.8 5.7 * time in minutes This indicates that orally administered iaeulin with lecithin alone has no effect on blood glucose 15 lowering. Example 3 A further experiment, not within the scope of the present invention, was performed for comparative purposes. 20 Oral insulin (100 units) was formulated with sodium salicylate and alkali metal edetate (both 5% by wt.) to evaluate its efficacy of blood glucose lowering in fasted state in healthy volunteers. voluiiceers were askeu~t;u 'GvciAxiylit- 25 have any breakfast prior to dosing. Volunteers were asked to take this oral insulin formulation in their mouth and swallow it. Blood glucose levels were monitored every 15 minutes using Bayer's glucometer Elite for 3 hours and the average results for 5 30 volunteers are shown in Table IV, AMENDED SHEET -is- Si- O : • 905 27S 768/-» +4-9 89 2 ^ ^ 5-02-2000 vuu "'L'L'" in'u,|^" - wv^'u IIU.uitu CA009900106 - 27 -Table IV Time*:0 15 30 45 60 75 SO 120 150 180 Avg; 5.8 5.8 5.B 5.9 5.8 5.9 5.7 5.9 6.2 6.0 * time in minutes 5 • This indicates that orally administered insulin with sodium salicylate and alkali metal edetate alone has no effect on blood glucose lowering. In addition, this formulation caused irritation and burning sensation, which lasted for several hours. 10 Example 4 A further experiment, not within the scope of the present invention, was performed for comparative purposes. Oral insulin (100 units) was formulated using 15 sodium salicylate and alkali metal edetate (both 5% by wt.) with Phospholipon-H (10 mg) and tested on healthy subjects. Blood glucose levels were monitored every 15 minutes using Bayer's glucometer Elite for 3 hours and the results are shown in Table V. 20 Table V Time*: 0 15 30 45 60 90 120 180 Avg: 5.3 5.3 5.3 5.4 5.6 5.7 5.7 5.8 * time in minutes _____ -^is"" indicates' thai:"orally administered ' insulin. 25 with sodium salicylate, alkali metal edetate and Phospholipon-H has no effect on blood glucose lowering. Example 5 Another experiment, not within the scope of the present invention was performed for comparative 3 0 purposes. Oral insulin (50 units) was formulated using only AMENDED SHEET — JliTvi-lltN :-2000 :15- 2- O : 22:59 umui i unn u wvut 905 276 7687-+ (-Ht9 89 2: qA Q09900106 - 28 - alkali metal lauryl sulphate (5% by wt) . Blood glucose levels were monitored every 15 minutes using Bayer's glucometer Elite for 3 hours and Che average results for four volunteers are shown in Table VI. S Table VI Time*s 0 IS 30 60 90 120 180 Avg: 5.8 5.6 5.4 5.3 5.4 5.4 5.6 * time in minutes This data shows that orally administered insulin 10 with only alkali metal lauryl sulphate has little metabolic effect on the blood glucose lowering in healthy subjects. This formulation caused substantial burning sensation and irritation in the subjects and lasted for two days. IS Example 6 Yet another experiment, within the scope of the present invention, was performed. Mixed micellar oral insulin (50 units) was formulated using alkali metal lauryl sulphate and sodium 2 0 salicylate (both 4.4% by wt.) and alkali metal edetate (2.2% by wt) with Phospholipon-H (10 mg) and tested on healthy volunteers. The method involved mixing the sodium lauryl sulphate, sodium salicylate ^d^ik^i metal~edeta.te 25 with water in a beaker with a magnetic stirrer at medium speed until the ingredients were dissolved, to form buffer solution. Insulin powder was placed in a beaker and to this powder was added the buffer solution. The solution was continuously stirred using a magnetic stir 3 0 bar until all of the insulin powder was dissolved and a clear solution obtained. The micellar solution so AMENDED SHEET L'tivcnr.M u^: : Lb- li- U : i23:00 : 905 27(3 76S7-> +4-y 89 2 ^ a nnnnAAHnc -02-2000"vv u iri"JL' vj.yiTu CA 009900106 - 29 - formed was stored in clean glass bottles and re f rigerated. Mixed micellar liposomal insulin was then prepared in a glass beaker, in which, was placed the Phospholipon-5 H and a small amount of isopropyl alcohol. The mixture was stirred at a high speed (1000 rpm) for about 10 minutes to ensure complete dissolution of the Phospholipon-H. To this solution was added the micellar insulin solution very slowly, drop wise, -using a glass 10 dropper, with continuous stirring at a high speed. The solution was stirred continuously for another 30 minutes at a high speed to ensure uniform micellar particle Bize distribution. Samples of the mixed micellar solution were taken 15 orally by the volunteers. Blood glucose levels were monitored every 15 minutes using Bayer's glucometer Elite for 3 hours and the average results for 5 volunteers are shown in Table VII. 20 Table VII Time*: 0 15 30 45 60 90 120 150 180 Avg: 6.5 6.1 5.5 5.3 5.3 5.4 5.5 5.5 5.5 * time in minutes __ "This""data""showsti^Coraxiy^adiainisrerea insul in ~ 25 with alkali metal lauryl sulphate combined with the sodium salicylate and alkali metal edetate with Phospholipon-H ha3 a small metabolic effect on blood glucose levels in healthy volunteers. Example 7 30 An experiment, within the scope of the present invention, was performed. In this example, the AMENDED SHEET - <jr.iv-iiui\« u- :15" 2" 0 : : 00 : 905 276 7687-» +49 89 2 r A nnQQnn 1 Ofi 15 02 2000 o-jui »• utsiui. jr.i u r.vuu hu-oitv UauuyyuuIUD - 30 - formulation was for oral administration. Oral insulin (50 units) was formulated using alkali metal lauryl sulphate and sodium salicylate (both 4.4% by wt.) and alkali metal edetate {2.2% by wt.) with 5 Phospholipon-H (10 mg) and sodium hyaluronate (1.1% by wt) . This formulation was tested on healthy subjects under fasting condition. The method involved mixing the sodium lauryl sulphate, sodium salicylate and alkali metal edetate 10 with water in a beaker with a magnetic stirrer at medium speed until the ingredients were dissolved, to form buffer solution. Insulin powder was placed in a beaker and to this powder was added che buffer solution. The solution was continuously stirred using a magnetic stir 15 bar until all of the insulin powder was dissolved and a clear solution obtained. The micellar solution so formed was stored in clean glass bottles and refrigerated. Mixed micellar liposomal insulin was then prepared 20 in a glass beaker, in which was placed the Phoepholipon-H and a small amount of isopropyl alcohol. The mixture was stirred at a high speed (1000 rpm) for about 10 minutes to ensure complete dissolution of the Phospholipon-H. To this solution was added the micellar 25 insulin solution very slowly, drop wise, using a glass dropper, with continuous stirring at a high speed. The solution was stirred continuously for another 30 minutes at a high speed to ensure uniform micellar particle size distribution. The hyaluronate and small amounts of 30 menthol and gortitol were then added, with continuous stirring. AMENDED SHEET A - - •<oijwrm-.i\ uj 2- 0 : 23:00 : 005 276 7687-- +49 89 £OA nnQQnnmfi 02 2000 u-u.iin wPim-iunn -J MWV/ITJ UAOuyyuuiuo - 31 - Samples of the mixed micellar solution were taken orally by the volunteers. Blood glucose levels were monitored every 15 minutes using Bayer's glucometer Elite for 3 hours and 5 the average results for 5 volunteers are shown in Table VIII. Table VIII Time*: 0 15 30 45 60 90 120 150 180 Avg: 6.5 5.9 5,6 5.4 4.9 5.0 4.9 5.2 5.4 10 * time in minutes This data shows that orally administered insulin with alkali metal lauryl sulphate,' sodium salicylate, alkali metal edetate, Phospholipon-H and sodium hyaluronate has resulted in lowering of blood glucose 15 levels in healthy subjects better than the above mentioned formulations. Example a A further experiment, within the scope of the present invention, was performed. In this example, the 20 formulation was for oral administration. A buffer solution was prepared using 0.5 g sodium lauryl sulphate, 0.5 g sodium salicylate and 0.25 g disodium edetare dissolved in 10 mL of water. The ^"^^luFi^~wa^a^ed^o^nsini^"a^~lnix^""to~foiOT^nI^ilar 25 insulin. Separately, 100 mg of powdered Phosphatidylcholin-H was added to a glass beaker and to this powder was added 10 mL 50% ethanol. The powder was dissolved completely. To this solution IS ftig (400 units) of micellar insulin 30 solution dissolved in 3 mL of the buffer solution to (give 30 units/mL insulin solution) was added slowly with vigorous mixing, to form a mixed micellar solution. AMENDED SHEET : J r>- 1!- u : Uli '■ 00 '■ GuS 27G 7687-» +49 89 •■> „ 15-02-2000'" "•*""«> -nnuiunn u mvwj -iwui-U CA 009900106 - 32 - To this was added 0.6 mL of sodium hyaluronate and 0.2 ml of 2% menthol solution containing 3% sorbitol. In one set of tests, ten Type II diabetic human volunteers who took insulin, by injection three times a 5 day, were studied. In another set of tests the volunteers were tested with insulin, taken orally. The volunteers fasted from midnight prior to the test, with no food being taken during the 4 hour study. On the first day, the volunteers received 10 units 10 insulin by injection {regular fast acting insulin, available from Eli Lilly) . On the second day, the volunteers received 30 units (1 mL volume per drop, approximately 20 drops) of the above-prepared oral insulin {3 times the injection dose). In both tests, 15 blood glucose levels were monitored every 15 minutes by Bayer's Glucometer Elite. The results, showing the average for the ten volunteers, were as shown on the following page: AMENDED SHEET "UfcNUHhN U2 :15- 2- 0 : 23:01 : 905 276 7bS7-> +49 89 2 o a nnoonnmc 15.Q2-2000""u -"-IIW. iutiv « i/wUJ .iw j ITU CA 009900 lUb - 33 - Table IX Blood glucose levels (mmol/L) Time (minutes) Oral Dose Injection (30 units) (10 units) 0 6.4 6.8 15 5.8 6.9 30 5.4 6.1 45 5.3 5.8 * 60 5.3 5.8 75 5.2 5.a 90 5.2 5.4 105 5.2 5.4 120 5.1 5.2 13S 5.1 5.1 150 5.2 4.9 165 5.3 4.9 130 5.3 4.8 195 5.4 4.8 210 5.4 5.2 225 5.6 5.2 240 5.6 5.4 The results show that the oral insulin fonnulation of the present invention, at a dosage of three times »*.. 4»U<* /»J 1 4 ¥>AVS1 a +*V| A WAAGUA VpUC -i-w » w-s*> *•-' ^ — w — — 25 injected insulin. Example 9 This example illustrates a method for making a mixed micellar fonnulation according to the present invention. 30 In a 250 mL capacity glass beaker was added 5 g AMENDED SHEET 15-02-2000 0'2 • 15- '2- 0 : 23-OJ : 005 276 76U7- +4-9 89 2:PA nnQQClOIDfi ■v vv.v ~j * •* t • !ii unmwvjnu u n«vw u • "i rj Un UU^yUU IUu - 34 - sodium lauryl sulphate, 5 g sodium salicylate and 2.5 g edetate. The beaker was placed on the hot plate with a magnetic stirrer. To this dry powder mixture was added 100 mL distilled water and the mixture was stirred, 5 using the magnetic stir bar, at a medium speed until all the powder was dissolved. The buffer solution was stored in a clean glass bottle at room temperature (pH 6.5). A micellar insulin solution was then prepared in a 10 50 mL capacity glass beaker, into which was placed 11.54 mg insulin powder. To this powder was added 10 mL of the buffer solution. The solution was continuously stirred using a magnetic stir bar until all of the insulin powder was dissolved and a clear solution 15 obtained. The micellar solution so formed was stored in clean glass bottles and refrigerated. A 2% menthol solution was then prepared from 100 mg menthol crystals, dissolved in 5 mL ethanol. To this solution was added 5 mg FD &. C blue dye. The solution 20 was stirred for 10 minutes and stored in a glass bottle at room temperature. Mixed micellar liposomal insulin was then prepared in a 50 mL glass beaker, in which was placed 100 mg of phosphat idylchorihe^TSigmaT"type^X=EH7^hyar^enacea;~~." Tcr 25 this powder was added 10 mL of isoprqpyl alcohol. The mixture was stirred at a high speed (1000 rpm) for about 10 minutes to ensure complete dissolution o£ the phosphatidylcholine. To this solution was added the micellar insulin solution very slowly, drop wise, using 30 glass dropper, with continuous stirring at a high speed. The solution was stirred continuously for another 3 0 AMENDED SHEET * - ui; • IS" U : 2y:01 : 305 276 7687-. +49 89 2;" . 15-02-2000 unuu.y^n u .vm CA009900106 - 35 - minutes at a high speed to ensure uniform micellar particle size distribution. To this solution was added 1 mL of the 2% menthol solution and 50 mg sodium hyaluronate. The semi-clear, translucent, light blue 5 colour, liposomal insulin mixed micellar solution (final volume 15 mL] was stored in a clean glass bottle and refrigerated. The solution had a pH of 6.5. -If the phosphatidylcholine powder does not dissolve completely, then heating up to about 45°C may be 10 required, e.g. using a water bath. It has been found that if the micellar insulin composition is not added slowly, then the mixed micellar formulation will not be formed and the formulation will be gelatinous and sticky. 15 Example 10 The formulation of Example 9 was tested in a manner similar to that indicated, in Example 8 except that the formulation of the present invention waa administered nasally. 20 On the first day, the ten volunteers each received 10 units insulin injection (regular fast acting, Eli Lilly) . On the second day, the volunteers received 20 units of the "oraln insulin of Example 9 (2 times the injection dosefjTT The-"orals~ xnsuxin~was™«iuiuj;rrj;oi-€:x£*-»~~as-25 drops (0.4 mL volume per drop, approximately 4 large drops in total, i.e. two drops in each nostril). The results, showing the average for the ten volunteers, are shown on the following page: 30 AMENDED SHEET 02-2000 - "UtNCHbN 0J. '• 15- 2- 0 : 23=01 : 905 276 7687-» +4-9 89 2 PA nnaannmfi |(-|C> i y u v -j'jt.m jnimiuTii u mvuu " OA UUyyUU IUO - 36 -Table 2 Blood glucose levels (mmol/L) Time (minutes) Nasal Dose Injection (20 units) (10 units) 0 7.4 6.8 15 6.7 7.0 30 5.9 £.8 45 5.3 6.3 60 5.0 6.3 75 5.2 5.8 90 5.1 5.2 105 5.0 5.0 120 4.6 5.2 135 4.5 4.2 150 4.3 4.6 165 4.3 4.0 180 4.8 4.1 195 5.3 4.3 210 5.4 4.5 225 5.7 4.7 240 5.6 5.0 The results show that the nasal insulin formulation of the present invention, at a, dosage of twice the injected level, is comparable to the injected insulin. 25 Example 11 The formula of Example 9 was taken and tests performed to determine the insulin action on meal glucose on healthy volunteers. Usually, diabetic patients take an insulin 3 0 injection 30 minutes prior to a meal, because injected insulin takes a long time to take effect. Injected. AMENDED SHEET -02-2000 'Jt-NCHLN <J2 : 15- 2- 0 '• 23 '02 '• 905 276 7887-» +4-9 89 2 /->A nnQQADinfi uuu u • jii H' unm-: vjrv\ u fivu-J ny-JiTU L»M Uu33w luu - 37 - insulin is slowly absorbed into bloodstream within 60 minutes and has metabolic effect on meal glucose levels. The mixed micellar formulation of Example 9 was tested in healthy volunteers under controlled conditions 5 to determine the oral insulin effect on meal glucose when compared to injected insulin. In one set of tests, ten healthy non-diabetic human volunteers were tested with insulin, by injection, in another set of tests the volunteers were tested with 10 insulin, taken orally. The volunteers fasted from midnight prior to the tests, with food being taken 30 minutes after dosing. The meals were standard Sastacal 240 mL liquid diet approved by the Diabetic Society, containing 400 calories. 15 On the first day, the volunteers received 10 units insulin by injection (regular fast acting insulin, available from Eli Lilly) . On the second day, the volunteers received 30 units of the above-prepared oral insulin (3 times the injection dose) . In both tests, 20 blood glucose levels were monitored every 15 minutes by Bayer's Glucometer Elite. The results are shown on the following page: AMENDED SHEET 15-02-2000 v*£ » xo- jL — v * ' yj^> • vjuu ^,/kj too* * nnnonn 1 nc v v g o * u j i iti unmiiunu u mvi uv n v • u i t u L//"\ UUyyUU I VJu - 38 - Table XI Blood glucose levels (mmol/L) Time (minutes) Oral Dose Injection (30 units) (10 units) 0 5,7 5.5 15 5,2 5.6 30 5.0 5.4 45 5.3 5.4 60 5.4 5.6 75 6.3 6.6 90 6.9 7,0 105 6.0 5.9 120 5.5 5.6 135 5.5 5.1 150 5.1 4.8 265 4.5 4.6 180 4.8 4.3 The results indicate that the oral insulin helps 20 control meal glucose levels in healthy volunteers when compared to injected insulin. Example 12 The mixed micellar formulation of Example 9 was tested in diabetic vollmtee^~uhlier^^troTIed 25 conditions to determine the oral insulin effect on meal glucose when compared to injected insulin. In one set of tests, ten Type II diabetic human volunteers who took insulin, by injection three times a day, were studied. In another set of tests the 30 volunteers w^re tested with insulin, taken orally. The volunteers fasted from midnight prior to the tests, with AMENDED SHEET 15-02-2000 JtiMv-titN vz • JS- 2- 0 : 23:02 : 905 270 7687-> +49 89 21 ~ . nnnnnninc ' u-'Juiiti uniiiuunu u iiiuou \u-sii-y GA009yOUIUb - 39 - food being taken 30 minutes after dosing. The meals were standard Sastacal 240 mL litjuid diet approved by the Diabetic Society, containing 400 calories. On the first day, the volunteers received 10 units 5 insulin by injection (regular fast acting insulin, available from Eli Lilly) . On the second day, the volunteers received 30 units of the above-prepared oral insulin (3 times the injection, dose) . In both tests, blood glucose levels were monitored every 15 minutes by 10 Bayer's Glucometer Elite. The average results for the 10 volunteers were as follows: Talale XII Blood glucose levels (imaol/L) Time (minutes) Oral Dose Inj ection (30 units) (10 units) 0 8.8 8.7 15 8.1 8.8 30 8.0 8.9 45 8.4 10.1 60 H O 10 11.8 75 11.8 11.8 90 12.3 12.2 10b 1U.S u.is 120 9.6 10.4 135 8.1 8.4 150 6.9 7.3 155 6.2 6.5 1B0 4.8 4.3 30 The results indicate that oral insulin helps to control meal glucose levels in diabetic patients when AMENDED SHEET \J V J • V tf I IB :35- 2- 0 ; 23:02 L>mwi J una u mvvy 905 276 7687-f ,,++9 8? 2: QA 0099001 06 - 40 - compared to injected insulin. Example 13 A chewable gum insulin fonnulation was prepared by vigorously stirring the liposomal insulin mixed micellar 5 solution of Example 9 while adding guar gum, beeswax, powdered acacia, oleic acid, gamma-linoleic acid and sorbitol, Por each 30 units of insulin, the mixture contained 100 mg guar gum, 50 mg beeswax, 50 mg powdered acacia, 100 mg oleic acid, 100 mg gamma-linoleic acid 10 and 1 mL 3% sorbitol in ethanol solution. The mixture was then poured into a flat tray cpated with polytetrafluoroethylene until the mixture was about 10 mm deep. The mixture then solidified and after solidification was cut into sticks about 1 cm by 3 cm. 15 Each stick contained about 30 units insulin. The mixed micellar formulation in chewable stick form was tested in diabetic volunteers under controlled conditions to determine the oral insulin effect on meal glucose when compared to injected insulin. 20 In one set of tests, five Type II diabetic human volunteers who took insulin, by injection three times a day, were studied. In another set of tests the volunteers were tested with the chewable gum insulin, __—oi" ally-.—Tie-voiunfcssss—£-asisd—Sres-^dnishfc—prio-E. 25 to the tests, with food being taken 30 minutes after dosing. The meals were standard Sastacal 240 mL liquid diet approved by the Diabetic Society, containing 400 calories. On the first day, the volunteers received 10 units 30 insulin by injection (regular fast acting insulin, available from Eli Lilly) . On the second day, the AMENDED SHEET — • • "'JEMr 02 15-02-2000 uJ -"VJM" 15- 2- 0 •• uni\u:vjrm u mvu1. 905 276 768" +49 89 2. iiy 1 u i T'J CA 009900106 - 41 - volunteers received 30 units of the above-prepared chewable gum oral insulin (3 times the injection dose). In both tests, blood glucose levels were monitored every 15 minutes by Bayer's Glucometer Elite. The average results for the five volunteers were as follows: Table XIII Blood glucose levels (mmol/L) 10 Time (minutes) Oral Dose Injection {30 units) (10 units) 0 9.1 8.8 15 9.3 8.2 30 9.3 8.0 45 10.2 8.4 60 11.2 9.2 75 12.1 10.3 90 12.9 11.8 105 13.2 11.6 120 12.8 11.0 135 12.2 10.2 150 11.6 9.6 165 11.0 9.5 180 10.6 9.1 A ^ is b 10.0 War ■ 210 9.5 8.2 225 8.8 8.0 240 8.2 7.5 Examnle 14 Another experiment, within the scope of the present 30 invention, was performed. In this example, the formulation was for oral administration. 1 AMENDED SHEET - - .'JV^V. H-iv iz- -J.- u : i:y:o3 : b05 276 7G87-» +49 89 2C „ . ^ 15-02-2000 uuu L"""'uni w .iuwitj CA009900106 - 42 - A buffer solution was prepared using 0.5 g sodium lauryl sulphate, 0.5 g sodium salicylate and 0.25 g disodium edetate dissolved in 10 mL of water. The solution was added to 8 mg (200 units) insulin and 5 mixed, to form micellar insulin. To this micellar solution were added 0.2 g bacitracin and 0.5 g evening primrose oil and the solution was mixed vigorously to form a oiixed micellar insulin solution (about 20 units/raL). 10 Six human volunteers were studied. The volunteers fasted from, midnight prior to the test, with no food being taken during the 4 hour study. On the first day, the volunteers received 10 units insulin by injection (regular fast acting insulin, 15 available from Eli Lilly) . On the second day, the volunteers received 20 units of the above-prepared oral insulin (twice the injection dose) . In both tests, blood glucose levels were monitored at intervals by Bayer's Glucometer Elite. 20 The results, showing the average for the six volunteers, were as follows: Table XIV Blood glucose levels (mmol/L) Time (minutes) Oral iiose" - AJl* J CiVrf liXUU 25 (20 units) (10 units) 0 8.8 7.9 15 8.4 7.9 30 8.1 8.2 45 7.4 8.3 30 60 6.3 7.6 90 5.1 6.2 AMENDED SHEET . . mju • ID- ^- u *. :03 : bOS 276 4-49 By 2v o A nnnnnnin^ «j«jv v'uvi.u uniMuunu u «vyg nuwnu Un UUyyUU IwO 15-02-2000 - 43 - 120 o in 5.2 150 4.8 4.6 180 5.1 3.9 210 5.3 4.4 240 5.6 5.2 The results show that the oral insulin formulation of the present invention, at a dosage of twice the injected level, is comparable to the injected insulin.. Example 15 10 A further experiment was performed to show another method of making the mixed micellar formulation of the present invention. In a 250 mL round bottom flask was added 100 mg of saturated lecithin powder {Phospholipon-9 OH} purchased 15 from the American Lecithin Co- To this powder was added 5 mL of absolute ethanol (I3SP grade) . The flask was tben attached to a rotary evaporator equipped with a vacuum pump and nitrogen inlet for inert atmosphere condition to minimize oxidation of the lecithin. The 20 flask was rotated at 100-150 rpm under vacuum. The solution in the flask was heated to 60°C by means of water bath to dissolve the powder completely. After complete dissolution of the powder, heating- was stopped arid the rotation speed was increased to 300 rpm, under 25 vacuum in nitrogen atmosphere until the alcohol evaporated completely, leaving a uniform film on the side of the flask. The rotation was continued for at least 30 minutes to ensure uniform coating of film on the wall and complete solvent removal. After 30 30 minutes the rotation was stopped and the vacuum was released. AMENDED SHEET </div>

Claims

<div class="application article clearfix printTableText" id="claims"> 5-02-2000 •L-iNN-ucv v-i ♦ J.o- ~.t- u ♦ 'J.ti 0*3 i 905 2/6 7GB7-* +4-9 89 v-s. ♦ xw- -i- u ♦ • c?vo «£ /o /t>©/— oy -i,: A nnnnriA-l nc uuitr unhiM jn-i u muy; iiv-vMtl OA (JUyyUU 1UO - 44 - To this flask was added micellar insulin solution which had been prepared from an aqueous solution of insulin, sodium lauryl sulphate, sodium salicylate and disodium edetate. The flask was shaken with the help of S shaker plate. Shaking was continued for at least 30 minutes and then the solution was sonicated with a high frequency sonicating probe for another SO minutes in order to form small uniform mixed micelles. The mixed micelles so obtained were analyzed by Malvern Zeta 10 (trade mark) particle size distribution measurement equipment equipped with the laser light scattering device. The mixed micelles particle size distribution obtained by this method was between 2 and 9nm. To this solution was added 1 mL of 2% menthol solution 1$ and 50 mg sodium hyaluronate. The semi-clear, translucent, light blue colour solution (final volume 10 mL) was stored in a clean glass bottle and refrigerated. The solution had a pH of 6.5. Example 16 2 0 Another experiment, within the scope of the present invention, was performed. K buffer solution was prepared using 0.5 g sodium lauryl sulphate, 0.5 g sodium salicylate and 0.25 g disodium edetate disi^oi^^~ih~rO"nffi"or~ waterTT -ine 25 solution was added to B mg <200 units) insulin and mixed, to form micellar insulin. To this micellar solution were added 0.5 g borage oil and the solution was mixed vigorously to form a mixed micellar insulin solution (about 20 units/mL) . AMENDED SHEET 15-02-2000 —-fcNCMJfcN OV :15- 2- 0 ; 20:03 : 905 276 7687- +49 89 25 ~ A rvnoonrn no ^p.uuu u ■ u u i 'ii i/niiu. jnu « ».VJU nu'Ui-ru CA 009900 106 - 45 - CLAIMS: 2,. A process for making a proteinic pharmaceutical formulation suitable for delivery through mucosal membranes comprising: 5 a) preparing a proteinic pharmaceutical agent composition in micellar form in an aqueous medium which has an alkali metal salicylate in a concentration of from 1 to 10 wt./wt.% of the total formulation, an alkali metal CS to C22 alkyl sulphate in a concentration 10 of from 1 to 10 wt./wt.% of the'total formulation acid a pharmaceutically acceptable edetate in a concentration of from 1 to 10 wt./wt.% of the total formulation; b) slowly adding the micellar proteinic pharmaceutical agent composition, while mixing, to at least one 15 absorption enhancing compound, while continuing to mix vigorously, said absorption enhancing compound being selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic 20 acicl, lactic acid, chamomile extract, cucumber extract, oleic acid, linolenic acid, borage oil, evening primrose oil, menthol, trihydroxy oxo cholanylglycine and pharmaceutically acceptable salts thereof, glycerin, polyglycerinT"lysine, poiyiysiiie7~polidDcaxioi^ alkyl —— 25 ethers and analogues thereof, triolein and mixtures thereof, wherein the amount of each absorption enhancing compound is present in a concentration of from 1 to 10 wt./wt.% of the total formulation, and the total concentration of alkali metal salicylate, alkali metal 3 0 C8 to C22 alkyl sulphate, edetate and absorption AMENDED SHEET i0M(_nr.v -IS- U : 2:d"-04- : 905 276 7687- +4:9 89 2:" A /-^ u u u u • u. i in uninu jnu u mv uu nv 1 J i 'U CA 0099001UO 15-02-2000 - 46 - j enhancing compounds is less than 50 wt./wt.% of the formulation. 2 . A process according to Claim 1 wherein there is an additional step of adding, while continuing mixing, at 5 least one absorption enhancing compound different to that added in step b), selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic acid, lactic 10 acid, chamomile extract, cucumber extract, oleic acid, linolenic acid, borage oil, evening primrose oil, trihydroxy oxo cholanylglycine, glycerin, polyglycerin. Lysine, polylysine, triolein and mixtures thereof. 3. A process according to Claim 1 wherein the 15 absorption enhancing compound in step b) is selected from the group consisting of saturated phospholipid, unsaturated phospholipid, phosphatidylcholine, phosphatidyl serine, sphingomyelin, phosphatidylethanolamine, cephalin, lecithin, 20 lysolecithin and mixtures thereof. 4. A process according to Claim 1 wherein one of the absorption enhancing compounds is lecithin and another absorption enhancing compound is selected from the group consistingofhyalurohic^acicT^pSarjrsceuticariy 25 acceptable salts of hyaluronic acid and mixtures thereof, the concentration of said another enhancing compound being from about 1 to about 5 wt./wt.%. 5. A process according to Claim 1 wherein the absorption enhancing compounds comprise combinations 30 selected from the group consisting of i) saturated AMENDED SHEET kt'£ • its** jL- O • ^3 • 04- » 90S 276 7(507—» +4«9 89 2« /—* a nnoonninfi v. v g • v - 47 - phospholipid and sodium hyaluronate; ii) saturated phospholipid and glycolic acid; iii) lecithin and sodium hyaluronate; iv) saturated phospholipid, glycolic ae-id and lactic acid; v) sodium hyaluronate and evening 5 primrose oil; vi) saturated phospholipid and bacitracin; vii) saturated phospholipid, sodium hyaluronate and bacitracin; and viii) sodium hyaluronate, oleic acid, and gamma linoleic acid. 6. A process according to Claim 1 wherein the 10 proteinic pharmaceutical agent is selected from the group consisting of insulin, heparin, so-called low molecular weight heparin, hirulog, hirugen, huridin, interferons, interleukins, cytokines, mono and polyclonal antibodies, ehemotherapeutic agents, 15 vaccines, glycoproteins, bacterial toxoids, hormones, calcitonins, insulin like growth factors (IGF), glucagon like peptides (GLP-l), large molecule antibiotics, protein based thrombolytic compounds, platelet inhibitors, DN&, KMA, gene therapeutics, antisense 20 oligonucleotides, opioids, narcotics, analgesics, NSAIDS, steroids, hypnotics, pharmaceutical preparations for the treatment of chronic pain and morphine. 7- A process according to Claim 1 wherein in step b) the micellar proteinic phax^ceuticar^agerLtr composition -25 is added to lecithin, with sonication, to form a mixed micellar formulation; and c) while continuing to mix, adding at least one absorption enhancing compound selected from the group consisting of hyaluronic acid, pharmaceutic ally 30 acceptable salts of hyaluronic acid, octylphenoxypoiyethoxyethanol, glycolic acid, lactic AMENDED SHEET 'Ui\cnt:s\ V U 'J ' U J 1 III ; L5- 2- 0 . 23:04- unnuiunu u HVJV S05 276 7687- ,,^9 89 2: CA 009900106 - 48 - acid, chamomile extract, cucumber extract, oleic acid, linolenic acid, borage oil, evening primrose oil, trihydroxy oxo cholanylglycine, glycerin, polyglycerin, lysine, polylysine, triolein and mixtures thereof; 5 wherein the amount of lecithin and the absorption enhancing compound are each present in a concentration of from 1 to 10 wt./wt.% of the total formulation, and the total concentration of alkali metal salicylate, alkali metal C8 to C22 alkyl sulphate, edetate and 10 absorption enhancing compounds is less than 50 wt. /wt. % of the formulation. 8. A process according to Claim 1 wherein the absorption enhancing compound is formed into a film prior to the addition of the micellar pharmaceutical 15 agent composition. 9. A process according Claim 1 wherein subsequent to the addition of the micellar pharmaceutical agent composition a second absorption enhancing compound is added, said second absorption enhancing compound being 20 different from the absorption enhancing compound previously used. 10. A process according to Claim l wherein a phenol selected from the group consisting of phenol, methyl pHenol™a5d*^ixtures™thered1f~"is™^disd~to" tfie"" mixed 25 micellar formulation and the resulting formulation placed in a container, and the container is subsequently charged with a propellant. 11. A process according to Claim 10 wherein the propellant is selected from the group consisting of 3 0 tetrafluoroethane, tetrafluoropropane, dimethylfluoropropane, heptafluoropropane, dimethyl AMENDED SHEET ' tNChtTM U2 -15- 2- 0 ; 23-04 : 905 276 7b87- +4:9 89 2S ~ . nnnnon* or- 15-02-2000'y IJ 'J'J'''"""m 1 urvi u mvu,j nu ■ v ITU CA 009900106 - 49 - ether, n-butane and isobutane. 12. A process according to Claim 1 wherein the pharmaceutical agent is insulin. 13. A process according to claim 11 wherein the 5 pharmaceutical agent is insulin. 14. A mixed micellar proteinic pharmaceutical formulation comprising a proteinic pharmaceutical agent in micellar form, water, an alkali metal C8 to C22 alkyl sulphate in a concentration of from 1 to 10 wt./vrt.% of 10 the total formulation, a pharmaceutically acceptable edetate in a concentration of from 1 to 10 wt./wt.% of the total formulation, at least one alkali metal salicylate in a concentration of from 1 to 10 wt./wt.% of the total fonnulation, and at least one absorption 15 enhancing compound, said absorption enhancing compound being selected from the group consisting of lecithin, hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, octylphenoxypolyethoxyethanol, glycolic acid, lactic acid, chamomile extract, cucumber extract, 20 oleic acid, linolenic acid, borage oil, evening primrose oil, menthol, trihydroxy oxo cholanylglycine and pharmaceutically acceptable salts thereof, glycerin, polyglycerin, lysine, poly lysine, polidocanol alkyl ethers and analogues thereot, trioxein luixturss- 25 thereof, wherein the amount of each absorption enhancing compound is present in a concentration, of from 1 to 10 wt./wt.% of the total formulation, and the total concentration of alkali metal salicylate, alkali metal C8 to C22 alkyl sulphate, ededate and absorption 30 enhancing compounds is less than 50 wt./wt.% of the formulation. AMENDED SHEET - 'bNCHtN 02 :15- 2- 0 : 23"-05 : 905 276 7687-» +49 89 2C~A nnnnnnmc
02-2000 w v univiwunu u wvvJ'J iwoitv OAUvjyyUUiUo - 50 - 15. A mixed micellar pharmaceutical formulation according to Claim 14 comprising lecithin. IS. A formulation according to Claim 14 wherein the alkali metal C8 to C22 alkyl sulphate is sodium lauryl 5 sulphate and the alkali metal salicylate is sodium salicylate. 17. A fonnulation according to Claim IS wherein, the lecithin ia selected from the group consisting of saturated phospholipid/ unsaturated phospholipid, 10 phosphatidylcholine, phosphatidyl serine, sphingomyelin, phosphatidyls thanolamine, cephalin, lysolecithin and mixtures thereof. 18. A formulation according to Claim 15 comprising a second absorption enhancing compound selected from the 15 group consisting of hyaluronic acid, pharmaceutically acceptable Baits of hyaluronic acid and mixtures thereof, the concentration of said second absorption enhancing compound being from about 1 to about 5 wt./wt.% of the total formulation. 20 19. A formulation according to Claim 14 comprising a combination selected from the group consisting of i} sodium lauryl sulphate, sodium salicylate, disodium edetate, saturated phospolipid and sodium hyaluronate; ii) sodium lauryl^sulphatei sodiiM' s^icrylatev uiaodiu^- 25 edetate, lecithin and sodium hyaluronate; iii) sodium lauryl sulphate, sodium salicylate, disodium edetate, sodium hyaluronate and evening primrose oil; iv) sodium lauryl sulphate, sodium salicylate, disodium edetate, saturated phospolipid and bacitracin; v) sodium lauryl 30 sulphate, sodium salicylate, disodium edetate, saturated phospolipid, sodium hyaluronate and bacitracin; vi) AMENDED SHEET r •uciM>-nn.i\ VZ • 15- 2- 0 '• 23 • 05 . 905 276 7687-» +49 89 2pft 0000001 flfi _^Q2 2QQQ"yuw ^ ' u v uniuuuiit u mvuu iiicvitj Un UU53UU i uu - 51 - sodium lauryl sulphate, sodium salicylate, disodium edetate, sodium hyaluronate, oleic acid and gamma linoleie acid; vii) sodium lauryl sulphate, sodium salicylate, disodium edetate, saturated phospholipid, 5 and glycolic acid; and viii) sodium lauryl sulphate, sodium solicylate, disodium edetate, saturated phospholipid, glycolic acid and lactic acid. 20. A formulation according to Claim 14 wherein the pharmaceutical agent is selected from the group ^ 10 consisting of insulin, heparin, so-called low molecular weight heparin, hirulog, hirugen, huridine, interferons, interleukins, cytokine, mono and polyclonal antibodies, chemotherapeutic agents, vaccines, glycoproteins, bacterial toxoids, hormones, calcitonins, insulin like 15 growth factors (IGF), glucagon like peptides (GLP-1), large molecule antibiotics, protein based thrombolytic compounds, platelet inhibitors, DNA, SWA, gene therapeutics antisense oligonucleotides, opioids, narcotics, analgesics, NSAIDS, steroids, hypnotics, 20 pharmaceutical preparations for the treatment of chronic pain and morphine. 21. A formulation according to Claim 14 wherein the pharmaceutical agent iB insulin. 22. A formulation according to Claim 21 in which the 25 absorption enhancing compounds are lecithin and a second absorption enhancing compound selected from the group consisting of hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid and mixtures thereof. 30 23. A formulation according to Claim 14 wherein the formulation additionally comprises a phenol selected AMENDED SHEET - 52 - from the group consisting o£ phenol, methyl phenol and. mixtures thereof. 24. A formulation according to Claim 23 wherein the formulation ie contained in an aerosol container and the 5 container is charged with a propellant. 25. A formulation according to Claim 24 wherein the propellent is selected from the group consisting of tetrafluoroethane, tetrafluoropropane, dimethylfluoropropane/ heptafluoropropane, dimethyl 10 ether, n-butane and isobutane. 26. The use of a proteinic pharmaceutical agent in the manufacture of a micellar formulation for delivering said proteinic agent to the buccal mocosa comprising spraying an effective amount of said proteinic agent to the buccal mucosa using a metered dose dispenser, while resisting substantial inhalation of said proteinic agent. 27. The use according to claim 26 wherein the proteinic pharmaceutical agent is as claimed in claim 6. 28. The use according to claim 27 wherein the proteinic pharmaceutical agent is insulin. INTELLECTUAL PROPERTY OFFICE OF N.Z. 0 7 DEC 2001 RECEIVED AMENDED SHEET </div>